Enterobacteriophage PhiX174

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Enterobacteriophage PhiX174
Bacteriophage Phi X 174 Electron micrograph.gif

Electron micrograph of phage ΦX174

Systematics
Classification : Viruses
Area : Monodnaviria
Empire : Sangervirae
Phylum : Phixviricota
Class : Malgrandaviricetes
Order : Petitviral
Family : Microviridae
Subfamily : Bullavirinae
Genre : Sinsheimer virus
Type : Enterobacteriophage PhiX174
Taxonomic characteristics
Genome : (+) ssDNA circular
Baltimore : Group 2
Symmetry : icosahedral
Cover : no
Scientific name
Escherichia virus phiX174
Short name
ΦX174
Left
NCBI  Taxonomy: 10847
ICTV Taxon History: 201853873
Structure of the capsid of ΦX174
Genome of the bacteriophage ΦX174 with its 11 genes

The Enterobacteriophage PhiX174 , officially Escherichia virus phiX174 , out of date Enterobacteria phage phiX174 , German also Bacteriophage Phi X 174 or Coliphage φX174, or kurzX174 for short, is a single-stranded DNA virus (ssDNA) of positive polarity that infects bacteria of the species Escherichia coli . It is the only species officially recognized by the International Committee on Taxonomy of Viruses (ICTV) in the virus genus Sinsheimerviru (obsolete Phix174microvirus , also called bacteriophage PhiX174 sensu lato or ΦX174 group), but there are other suggestions (' Phage MED1 ').

Research history

The genome of ΦX174 is the first DNA-based genome to be fully sequenced, this work was completed by Fred Sanger and his team in 1977. Walter Fiers and Robert Sinsheimer had already shown in 1962 that the ΦX174 DNA is closed in a ring. In 1967, Nobel laureate Arthur Kornberg demonstrated on the ΦX174 as a model for the first time that DNA synthesized by purified enzymes in a test tube can generate all the characteristics of a natural virus, which ushered in the age of synthetic biology . In 1972–1974, Jerard Hurwitz , Sue Wickner and Reed Wickner, together with colleagues, identified the genes required to produce the enzymes that catalyze the conversion of the single-stranded form of the virus into the double-stranded, replicative form .

In 2003, Craig Venter's group reported that they were the first to assemble a viral genome - that of ΦX174 - entirely in vitro from synthesized oligonucleotides. The virus particle (virion) of ΦX174 was also successfully assembled in vitro . Recently it was shown how the strongly overlapping genome can be completely decompressed and still remain functional.

Genome

ΦX174 has a circular single-stranded DNA (ssDNA) of positive polarity . The genome consists of 5386 nucleotides that encode 11 proteins . Of these eleven genes, only eight are essential for viral morphogenesis . The GC content is 44% and 95% of the nucleotides belong to coding genes .

Mode of action

Infection begins when the G-protein binds to lipopolysaccharides on the surface of the bacterial host cell. As a DNA pilot protein, the H protein controls the viral genome through the bacterial membrane of the coli bacteria (Jazwinski et al. 1975) very probably via an assumed 'N-terminal transmembrane helix' (Tusnády, Simon 2001). However, H-protein has been shown to be a multifunctional protein (Cherwa, Young, Fane 2011). It is the only capsid protein of ΦX174 that is not in the crystal structure. It is low in aromatic amino acids and high in glycine , which makes the protein structure very flexible. In addition, at high concentrations (i.e. at the end of the virus's phase of replication), the H protein induces lysis (destruction) of the bacterial host, presumably because the assumed N-terminal transmembrane helix drills holes through the bacterial wall . In addition, Ruboyianes et al. Established in 2009 that the H protein is required for optimal synthesis of other viral proteins. Mutations in the H protein that prevent virus incorporation can be overcome if protein B (the internal scaffold protein) is supplied in excess.

Remarks

ΦX174 is regularly used as a positive control in DNA sequencing due to its relatively small genome size compared to other organisms, its relatively balanced nucleotide content - about 23% guanine , 22% cytosine , 24% adenine and 31% thymine , i.e. H. 45% G + C and 55% A + T .

See also

literature

  • PD Baas, GP van Heusden, JM Vereijken, PJ Weisbeek, HS Jansz: Cleavage map of bacteriophage phiX174 RF DNA by restriction enzymes . In: Nucleic Acids Res. , 3 (8), August 1976, pp. 1947-1960, PMC 343051 (free full text), PMID 1085927

Web links

Individual evidence

  1. ↑ a b c d e ICTV: ICTV Taxonomy history: Escherichia virus phiX174 , EC 51, Berlin, Germany, July 2019; Email ratification March 2020 (MSL # 35)
  2. ↑ a b c Coliphage phi-X174, complete genomeNCBI Reference Sequence: NC_001422.1 , NCBI
  3. ↑ Simon J. Labrie, Marie-Ève ​​Dupuis, Denise M. Tremblay, Pier-Luc Plante, Jacques Corbeil, Sylvain Moineau: A New Microviridae Phage Isolated from a Failed Biotechnological Process Driven by Escherichia coli (PDF) in: Journals ASM: Applied and Environmental Microbiology (AEM) Volume 80 No. 22, November 2014, pp. 6992-7000
  4. ↑ F. Sanger, GM Air, BG Barrell, NL Brown, AR Coulson, JC Fiddes, CA Hutchison, PM Slocombe, M. Smith: Nucleotide sequence of bacteriophage ΦX174 DNA . In: Nature . 265, No. 5596, 1977, pp. 687-695. bibcode : 1977Natur.265..687S . doi : 10.1038 / 265687a0 . PMID 870828 .
  5. ^ Walter Fiers, Robert L. Sinsheimer: The structure of the DNA of bacteriophage ΦX174 . In: Journal of Molecular Biology . 5, No. 4, 1962, p. 424. doi : 10.1016 / S0022-2836 (62) 80031-X .
  6. ^ The Arthur Kornberg Papers. "Creating Life in the Test Tube," in: National Library of Medicine Profiles in Science, 1959-1970
  7. ↑ Mehran Goulian, Arthur Kornberg, Robert L. Sinsheimer: Enzymatic Synthesis of DNA, XXIV. Synthesis of Infectious Phage ΦX174 DNA . In: Proceedings of the National Academy of Sciences . 58, No. 6, 1967, pp. 2321-2328. bibcode : 1967PNAS ... 58.2321G . doi : 10.1073 / pnas.58.6.2321 . PMID 4873588 . PMC 223838 (free full text).
  8. ^ Sue Wickner, Jerard Hurwitz: Conversion of X174 Viral DNA to Double-Stranded Form by Purified Escherichia coli Proteins , in: Proc Natl Acad Sci USA 71 (10), pp. 4122-4124, November 1974, doi: 10.1073 / pnas. 71.10.4120 , PMID 4610569 , PMC 434340 (free full text)
  9. ↑ Hamilton O. Smith, Clyde A. Hutchison, Cynthia Pfannkoch, J. Craig Venter: Generating a Synthetic Genome by Whole Genome Assembly: ΦX174 Bacteriophage from Synthetic Oligonucleotides . In: Proceedings of the National Academy of Sciences . 100, No. 26, 2003, pp. 15440-15445. bibcode : 2003PNAS..10015440S . doi : 10.1073 / pnas.2237126100 . PMID 14657399 . PMC 307586 (free full text).
  10. ↑ James E. Cherwa, Lindsey J. Organtini, Robert E. Ashley, Susan L. Hafenstein, Bentley A. Fane: In Vitro Assembly of the ΦX174 Procapsid from External Scaffolding Protein Oligomers and Early Pentameric Assembly Intermediates . In: Journal of Molecular Biology . 412, No. 3, 2011, pp. 387-396. doi : 10.1016 / j.jmb.2011.07.070 . PMID 21840317 .
  11. ^ Paul R. Jaschke, Erica K. Lieberman, Jon Rodriguez, Adrian Sierra, Drew Endy: A fully decompressed synthetic bacteriophage ΦX174 genome assembled and archived in yeast . In: Virology . 434, No. 2, 2012, pp. 278-84. doi : 10.1016 / j.virol.2012.09.020 . PMID 23079106 .
  12. ↑ GE Tusnády, I. Simon: The HMMTOP transmembrane topology prediction server . In: Bioinformatics , 17 (9), September 2001, pp. 849-850, PMID 11590105
  13. ↑ JE Cherwa Jr, LN Young, BA Fane: Uncoupling the functions of a multifunctional protein: the isolation of a DNA pilot protein mutant that affects particle morphogenesis . In: Virology , 411 (1), March 1, 2011 (online January 11, 2011), pp. 9-14, doi: 10.1016 / j.virol.2010.12.026 , PMID 21227478
  14. ↑ Mark V. Ruboyianes, Min Chen, Mathew S. Dubrava, James E. Cherwa Jr., Bentley A. Fane: The Expression of N-Terminal Deletion DNA Pilot Proteins Inhibits the Early Stages of φX174 Replication . In: J Virol. , 83 (19), October 2009, pp. 9952-9956, doi: 10.1128 / JVI.01077-09 , PMC 2748053 (free full text), PMID 19640994